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J A Ellis

Publications and source records attributed to J A Ellis.

142 records · Page 8Linked to original sources

Immunophenotype of pulmonary cellular infiltrates in sheep with visceral caseous lymphadenitis.

Pulmonary lesions associated with Corynebacterium ovis were analyzed with an indirect immunoperoxidase staining technique using monoclonal antibodies. The predominant cells in abscess walls and surrounding lung parenchyma were large macrophages which expressed major histocompatibility complex (MHC) class II molecules on their surfaces. T lymphocytes were prominent in the same sites in the naturally occurring lesions, and SBU-T4-positive ("helper/inducer") cells were the major subset of lymphocytes (mean T4/T8 ratio = 3.5). B lymphocytes and granulocytes comprised minor populations of infiltrating cells. These results implicate activated macrophages and MHC class II-restricted T lymphocytes in the pathogenesis of established C. ovis infections in sheep.

Animals↗

Monoclonal antibodies which react with bovine T-lymphocyte antigens and induce blastogenesis: tissue distribution and functional characteristics of the target antigens.

In this study we report on the tissue distribution and functional characteristics of bovine T-cell differentiation antigens recognized by the monoclonal antibodies (mAbs) IL-A26, IL-A27, and IL-A28. All three mAbs are able to stimulate proliferation of peripheral blood mononuclear cells (PBM) and inhibit proliferation of responder cells in mixed leucocyte cultures (MLC). MAbs IL-A27 and IL-A28 are believed to react with the same molecule, which is different to that recognized by IL-A26 as determined by a number of criteria. MAbs IL-A27 and IL-A28 inhibit binding of one another, but not of IL-A26. MAbs IL-A27 and IL-A28 react with 25% of thymocytes confined to the medulla, whereas IL-A26 reacts with approximately 80% of thymocytes, including medullary and cortical populations. MAbs IL-A27 and IL-A28 react with thymocytes which express BoT4 or BoT8 singularly, whereas IL-A26 reacts with all cells which express BoT4 or BoT8, either singularly or dually, in addition to all thymocytes which react with IL-A27/28. Only IL-A26 inhibits spontaneous sheep erythrocyte (E)-rosette formation by bovine T cells. Based on tissue distribution and functional characteristics, IL-A26 is believed to recognize the bovine homologue of CD2, designated BoT2, whereas IL-A27/28 reacts with a mature T-cell antigen. Cells reactive with the mAbs constitute approximately 60% of bovine PBM. Using these mAbs in dual immunofluorescence analyses, at least three populations of bovine T cells are demonstrable in PBM. The majority of T cells are BoT4+ or BoT8+ and also react with IL-A26/27/28. A second small population of PBM is negative for BoT4 and BoT8 but is IL-A26/27/28+. A third population (less than 5%) is BoT4-/BoT8-/ILA27/28- but reacts with IL-A26.

Animals↗

Bovine pan T-cell monoclonal antibodies reactive with a molecule similar to CD2.

Monoclonal antibodies (mAb) CH128A and CH61A react with molecules of 50,000-60,000 MW. They are expressed by all T cells in cattle, comprising 44-69% of peripheral blood mononuclear leucocytes (PBM), the majority of lymphocytes in T-dependent areas of lymph node, and 75-80% of cells derived from the thymus including both cortical and medullary thymocytes. The molecule recognized by these mAbs is not expressed on B lymphocytes, monocytes/macrophages, or granulocytes. Both mAb inhibit spontaneous rosette formation by sheep erythrocytes and bovine lymphocytes. We postulate that these mAb see the bovine homologue of the human sheep red blood cell receptor CD2 and has been named BoT2.

Animals↗

Bovine mononuclear phagocytic cells: identification by monoclonal antibodies and analysis of functional properties.

Monoclonal antibodies (mAb) which react with bovine monocytes have been produced. These include three mAb (P8, IL-A22 and IL-A24) that recognize the majority of monocytes and granulocytes in peripheral blood; two of these mAb were also shown to react with 30-40% of cells in bone marrow, including both monocytic and granulocytic cells, and with variable percentages of tissue macrophages. Thus these mAb can act as markers for myeloid cells in haemopoietic tissues and for monocytes in cell populations devoid of granulocytes. A further two mAb (IL-A23 and IL-A25) recognize monocytes and/or macrophages. The reactivity of one of these mAb (IL-A25) appears to be mainly restricted to pulmonary macrophages. The other mAb reacts with a variable proportion of blood monocytes and generally with a higher percentage of tissue macrophages, suggesting that its expression may relate to activation or maturation of monocytes. In order to study the functional properties of peripheral blood monocytes, techniques were developed for obtaining populations of peripheral blood mononuclear cells (PBM) depleted of monocytes to less than 0.2% and monocyte populations of greater than 97% purity. Removal of monocytes from PBM abrogated the capacity of the cells to proliferate in response to Con A and PBS, although addition of 2-mercaptoethanol to the cultures restored proliferation. In both allogeneic and autologous mixed leukocyte cultures (MLC), monocytes were required in the stimulator cell populations for induction of the proliferative responses, and both responses could be elicited with purified monocytes. However, proliferation in the autologous MLC occurred only with responder cell populations that were depleted of monocytes. Moreover, it was shown that addition of more than 5% unirradiated monocytes to the autologous MLC suppressed proliferation. These findings indicate that monocytes play an important role in the induction and regulation of cellular immune responses in cattle. Two of the mAb that react with monocytes and granulocytes were tested for their capacity to inhibit proliferative responses of PBM to mitogens, alloantigens or the soluble antigen, KLH.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Peripheral blood leucocytes subpopulation dynamics during Trypanosoma congolense infection in Boran and N'Dama cattle: an analysis using monoclonal antibodies and flow cytometry.

A panel of monoclonal antibodies (MoAb) with specificities for bovine leucocyte subsets were used in conjunction with routine haematological procedures to analyse sequential changes in peripheral blood leucocyte populations during the course of tsetse fly-transmitted Trypanosoma congolense infection in trypanotolerant N'Dama and trypanosusceptible Boran cattle. Subsequent to the first parasitaemic wave, the N'Dama cattle maintained packed cell volumes (PCV) above 22 and lower levels of parasitaemia than Boran throughout the 160 days of the experiment. In contrast, the Borans developed severe anaemia and required curative drug therapy (i.e., PCV dropped to less than 15) by 55 days (range: 22-55 days) post infection. There were significant (P less than 0.05) decreases in total white blood cells and total lymphocytes from pre-infection levels to the first peak of parasitaemia (day 16 post-infection) in both groups. Flow cytometric analyses using MoABs revealed that this change was due to an absolute decrease in T cells expressing BoT2 and either BoT4 or BoT8, surface immunoglobulin M-positive (sIgM+) B cells, and null cells which did not express T cell, B cell or monocyte markers. During this period there was significant variation over time, but no overall increase or decrease, in the number of cells expressing class II major histocompatibility (MHC) molecules or monocyte markers, or in the number of circulating neutrophils or eosinophils. The BoT4/BoT8 ratios were significantly (P less than 0.01) increased in both groups of infected animals at the first peak of parasitaemia. After day 22 in the infected N'Damas and in the Borans which required drug therapy, there was a leucocytotic response characterized by an increase in the total number of B cells, T cells, and null cells. Prior to infection and throughout the course of the experiment N'Dama cattle had significantly (P less than 0.01) higher numbers of B cells and null cells than Boran.

Animals↗

The properties and functional activity of non-lymphoid cells from bovine afferent (peripheral) lymph.

Following prescapular lymphadenectomy, the cellular composition of afferent (peripheral) lymph from eight out of 13 calves in which the afferent and efferent lymphatic ducts had anastomosed was examined using monoclonal antibodies (mAb) specific for bovine lymphoid populations. Afferent lymph leucocytes (ALL) consisted of around 50% T lymphocytes, of which 35% and 14% expressed BoT4 and BoT8, respectively, 20-25% B lymphocytes and 23% non-lymphoid cells. When populations of non-lymphoid cells were prepared from ALL by adherence to gelatin-plasma-coated flasks, 81% contained alpha-napthyl acetate esterase (ANAE) and 70% were phagocytic, whereas the non-adherent population contained around 5% of these cell types, the cells being largely non-phagocytic and negative for ANAE. Following purification of non-lymphoid cells on the fluorescence-activated cell sorter (FACS) using IL-A22 (mAb detecting macrophages and polymorphonuclear leucocytes, PMN), only 72% of the positive cells contained ANAE, 19% were phagocytic, and 84% expressed Fc1 receptors. While the ALL depleted of IL-A22+ cells could not proliferate in the presence of specific antigen in culture, the addition of purified monocytes from peripheral blood leucocytes (PBL) or IL-A22-positive ALL to macrophage-depleted PBL or IL-A22-negative ALL promoted proliferative responses to specific antigens. Similar populations of non-lymphoid cells stimulated mixed leucocyte reactions (MLR) in allogeneic combinations. When these non-lymphoid cells were pulsed with specific antigens prior to addition to autologous PBL or ALL in culture, normal or gamma-irradiated but not glutaraldehyde-fixed cells from PBL or ALL induced proliferation and immunoglobulin (Ig) synthesis in responder cells. Proliferation was depressed in cultures containing mAb detecting BoT4, MHC class II antigens and a mAb which detected non-lymphoid cells in PBL; Ig synthesis was inhibited only by anti-class II mAb. Monocyte-depleted PBL or IL-A22-negative ALL responded specifically to autologous antigen-presenting cells which had been pulsed with the original immunogen and not other antigens.

Animals↗

Alloreactive bovine T lymphocyte clones: an analysis of function, phenotype, and specificity.

A bovine alloreactive cell population was subjected to complement-dependent lysis with monoclonal antibody (mAb) IL-A11. The original population and the population depleted of cells bearing the determinant recognized by mAb IL-A11 were cloned. Parent cultures and 21 clones were examined for cytolytic function and for expression of determinants recognized by mAb IL-A11 and two additional mAb, IL-A12 and IL-A17. Clones could be classified according to maximal achievable levels of cytolysis by using Theileria parva-infected bovine lymphoblastoid target cells. In this way, three groups were identified--one capable of high level cytolysis, one of intermediate levels, and one group comprising apparently noncytolytic clones. The clones in the first group reacted with mAb IL-A17; those in the second and third groups, with mAb IL-A11 and IL-A12. It was shown that cytotoxicity effected by IL-A17+ clones could be inhibited by this mAb and also by a mAb directed to MHC class I determinants on target cells. Conversely, cytotoxicity effected by IL-A11+/IL-A12+ clones could be inhibited by mAb IL-A11 and by a mAb directed to MHC class II determinants on target cells. The levels of expression of class I and class II determinants on target cells correlated with the levels of killing by clones of the IL-A17+ phenotype and clones of the IL-A11+/IL-A12+ phenotype, respectively. The results indicate that cytotoxic bovine T lymphocyte clones specific for class I MHC antigens and both cytotoxic and noncytotoxic clones specific for class II MHC antigens can be obtained. Further, their specificity for class I or class II antigens can be determined by phenotyping with mAb.

Animals↗

Lesions and saliva-specific antibody responses in rabbits with immediate and delayed hypersensitivity reactions to the bites of Glossina morsitans centralis.

Rabbits exposed to feeding tsetse flies developed cutaneous hypersensitivity responses to fly bites. These responses had characteristics of immediate and delayed type hypersensitivity. Saliva components from the tsetse fly Glossina morsitans centralis were electrophoretically separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Major salivary proteins of 160, 92, 66, 64, 55, 42, 33, 28, and 15 kilodaltons were identified. Separated salivary components were transferred to nitrocellulose filters and probed with lectins and with whole sera and purified IgG from rabbits which had been exposed, via fly feeding, to tsetse antigens for variable periods. Many of the salivary proteins were identified as glycoproteins. Several major salivary proteins were recognized by antibodies from sensitized rabbits.

Animals↗

Characterization by a monoclonal antibody and functional analysis of a subset of bovine T lymphocytes that express BoT8, a molecule analogous to human CD8.

Monoclonal antibody (mAb) IL-A17 characterizes a subset of bovine T lymphocytes. IL-A17 recognizes a 34,000-35,000 MW doublet, designated BoT8, which is expressed on the surface of approximately 20% of peripheral blood mononuclear leucocytes (PBM), a subpopulation of lymphocytes in T-dependent areas of lymph nodes and spleen, and about 70% of thymocytes. This molecule is not expressed on B lymphocytes, monocytes/macrophages or granulocytes. Double labelling of PBM showed that the BoT8+ population is distinct from the T lymphocyte subset expressing BoT4. BoT8+ lymphocytes purified with a fluorescence-activated cell sorter (FACS) proliferated poorly in response to mitogenic and alloantigenic stimulation in the absence of exogenous growth factors. IL-A17 had no inhibitory effect on proliferation of PBM to mitogens (Con A and PHA) or alloantigens and no measurable effect on the in vitro generation of cytolytic effector cells. However, in some experiments IL-A17 was found to block partially allospecific cytolytic function mediated by bulk effector cell populations when included in 51Cr-release assays. Fractionation of effector cells generated in an allogeneic mixed leucocyte culture (MLC) demonstrated that cytotoxic cells specific for class I major histocompatibility complex (MHC) antigens reside within the BoT8+ population. Based on these data, and information reported elsewhere on alloreactive bovine T-cell clones, BoT8 is considered to be analogous to CD8 in humans and equivalent molecules in other species.

Animals↗

Ovine interleukin-2: partial purification and assay in normal sheep and sheep with ovine progressive pneumonia.

Interleukin-2 (IL-2), a T cell derived lymphokine, acts in nonspecific hormone-like fashion to maintain proliferation of activated lymphocytes in vitro and is believed to play a key role in cell-mediated immune function in vivo. The parameters of induction and assay of factors with IL-2 activity were examined in a group of clinically normal sheep seronegative for antibodies to ovine progressive pneumonia virus (OPPV-). Supernatants from cultures of Concanavalin A (Con A) stimulated mononuclear leukocytes (ML) derived from peripheral blood and lymph nodes contained factors with the capacity to maintain continued proliferation in Con A stimulated lymphoblasts. This activity was localized by gel chromatography to fractions containing proteins of 17,000-20,000 daltons. In a group of sheep seropositive for antibodies to OPPV (OPPV+), decreased levels of IL-2 activity were found in ML culture supernatants derived from the posterior mediastinal lymph nodes of sheep with clinical and pathological evidence of OPP, when compared to OPPV+ sheep with no lesions and sheep with visceral caseous lymphadenitis. This decrease in IL-2 activity appeared not to be associated directly with levels of prostaglandin E2 in these supernatants. These findings may correlate with virus induced alterations in cell mediated immune function in lymphoproliferative lesions of OPP.

Animals↗

Evidence of decreased concanavalin A induced suppressor cell activity in the peripheral blood and pulmonary lymph nodes of sheep with ovine progressive pneumonia.

Concanavalin A (Con A) induced suppressor cell activity was evaluated in a group of ovine progressive pneumonia virus-antibody positive sheep (OPPV+). Decreased levels of suppressor activity were observed in the peripheral blood and regional pulmonary lymph nodes of animals with clinical and pathological evidence of ovine progressive pneumonia (OPP) when compared to animals with no lesions and animals with caseous lymphadenitis involving the lungs and pulmonary lymph nodes. Decreased levels of Con A stimulated lymphocyte proliferation was also observed in the peripheral blood and iliac lymph nodes of sheep with OPP. Sheep with OPP were found to be hypogammaglobulinemic. Sera from OPPV+ sheep had no effect on T and B cell mitogen stimulated responses of lymphocytes from sheep seronegative for antibodies to ovine progressive pneumonia virus (OPPV-) when compared to normal sheep serum.

Animals↗

Immunomorphologic and morphometric changes in pulmonary lymph nodes of sheep with progressive pneumonia.

Morphologic, immunohistochemical, and morphometric studies were conducted on the posterior mediastinal lymph nodes of eleven sheep with naturally occurring ovine progressive pneumonia and four apparently healthy sheep with no pulmonary lesions (three seropositive, one seronegative for antibody to ovine progressive pneumonia virus). Compared with lesion-free sheep, sheep with ovine progressive pneumonia had a seven-fold increase in B lymphocyte areas and a 21/2-fold increase in T lymphocyte areas of these lymph nodes. Immunochemistry revealed cytoplasmic immunoglobulin G in scattered cells of germinal centers, medullary cords and interfollicular areas and membrane-associated immunoglobulin G in dendritic cells of germinal centers. Immunoglobulin M staining cells were widely scattered in germinal centers and medullary cords. Although B cell hyperplasia seemed to be the predominant process in lymph nodes of sheep with ovine progressive pneumonia, this was not accompanied by the expected degree of plasmacytosis, morphologically and immunohistochemically. These findings may represent an aberrancy of immunoregulation in ovine progressive pneumonia.

Animals↗

Ovine concanavalin A-induced suppressor cells: generation, assay, age-related effects and re-evaluation of mechanism of suppression.

Preactivation of ovine peripheral blood and lymph node mononuclear cells with mitogenic doses of concanavalin A (Con A) induced cells which suppressed mitogen-stimulated proliferative responses of untreated autologous and allogeneic responder cells. The degree of suppression varied with preactivating doses of Con A, length of preactivation time, and ratio of preactivated to responder cells. The role of macrophages in generation of suppressor cells was not evaluated. However, macrophages were not required to mediate suppression in cocultures, as lymphoblasts depleted of macrophage by plastic adherence and nylon wool columns mediated equal, and often greater, suppression than unseparated, preactivated cells. Suppressor cell activity in peripheral blood increased from the neonatal period to adulthood. Supernatants from Con A preactivated cell cultures with detectable interleukin-2 activity abrogated suppression when added at 0 and 24 hr of the 72 hr coculture period, suggesting that Con A-induced suppressor cells exert their function by decreasing available levels of IL-2 in the cocultures.

Aging↗

Injuries associated with in-line skating from the Canadian hospitals injury reporting and prevention program database.

In-line skating, also known as rollerblading, is an increasingly popular recreational activity that carries with it the potential for injury. As reported in the Canadian Hospitals Injury Reporting and Prevention Program database (CHIRPP), 194 children were injured while in-line skating. Fractures to the radius and ulna were the most common type of injury sustained (57.5%), followed by lacerations and abrasions (14.9%). Five children had concussions and very few children reported wearing protective gear such as a helmet or wrist, elbow and knee protectors. Compared to the database overall, in-line skaters suffered more severe injuries and were more likely to require follow-up treatment. Safety implications in relation to protective gear and learning the sport of in-line skating are discussed.

Adolescent↗